JPS6020394B2 - Antibiotic manufacturing method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides the antibiotic 7-[D-Q-ami/1Q-1(
p-hydroxyphenyl)acetamide]-3-(1,
The present invention relates to a method for purifying 2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid by converting it into a crystalline solvate that is highly suitable for oral or subcutaneous administration.

7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid is a novel cephalosporin. , also known as BL-S640, has been patented by our collaborators Davia Will and Leo MrdBmceCrast (U.S. S. 318340.1972
．． 12.26 application).

The specification states that 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]131 (1,2,3-
triazol-5-ylthiomethyl)-3-cephem-
The isolation of 4-carboxylic acid methanolates is described. There are two problems in the practical application of this compound.

Part 1 is 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid necessary for human use. The second problem is that it does not lose its vitality during storage, does not impair crystallinity, dispersion uniformity, etc., and also prevents oily, agglomerated, sedimentary, sticky, etc. It is a matter of formulating an aqueous dispersion suitable for oral and subcutaneous administration without undesirable changes. These problems are compounded by the fact that the compound loses its thiol component and decomposes very quickly at pH 7.0 or higher. Furthermore, crude products synthesized by chemical reactions are considerably contaminated by raw material residues and various decomposition products that cannot be reasonably removed by recrystallization, solvent washing, or the like. Attempts to crystallize and purify the zwitterionic product or hydrate have been unsuccessful.

Attempts to remove the solvent from the solvate to obtain an anhydrous product have also been unsuccessful, and the resulting product is sticky. Methanol is unsuitable for use by humans who are aware that methanol is toxic, and furthermore, the purification of meth/late does not proceed smoothly as measured by viability, decolorization, and reduction in impurity content.

Ethanolate was also prepared, but even this compound could not achieve the objective. The above compound crystallized but no purification could be achieved. Furthermore, when ethanolail was suspended in water, it gradually lost its ethanol, became a solid sticky substance, lost its crystallinity, and turned into a rubber-like substance. The object of the present invention is to obtain crystalline 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-13-cephemu-4-carvone. 1,2-propylene glycolate of the acid, preferably 1.0 to 1.6 moles of 1,2 per mole of the above carboxylic acid
- glycolate to which propylene glycol is attached, more preferably 1 mole of 1,
This is accomplished by preparing a glycolate with 2-propylene glycol attached.

The present invention also provides crystalline 71 [D-Q-amino-Q1 (
p-hydroxyphenyl)acetamide]-3-(1,
Also provided is a method for preparing 2,3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid 1,2-propylene glycolate. The method is to prepare 7-[D-ha-amino-Q-(p-hydroxyphenyl)acetamide]-13-(1,2,3-triazol-5-ylthiomethyl)-13-cephemu-4-carboxylic acid at a pH of about 2.0 or less. The acidic solution is raised in aqueous 1,2-propylene glycol to raise its pH value above 4.0, preferably from 4.0 to 5.0, by reduction of the base, thereby precipitating the crystalline propylene glycolate product, and then It can be recovered by a conventional method such as passing through a furnace or centrifuging. 7-[D-Q-ami/-Q-(p
-hydroxyphenyl)acetamide]-3-(1,2
, 3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid, the zwitterion of cephalosporin or its hydrate or solvate and
.
A solution of cephalosporanic acid is thereby produced.

The most preferred form of cephalosporin for this method is the methanolate. The volume of the solution is then raised by adding a sufficient amount of base, thereby crystallizing the propylene glycolate product.

Most preferably, the method uses a base at pH. This is a method in which impurities are precipitated from the solution by gradually adding it to a solution with a pH value of about 1.7 or less. The reaction mixture is optionally (but preferably) treated with carbon, and then the persistent products are removed by filtration. For acidic solutions, adjust the pH to 4.0 or higher by adding a base.
Preferably it is between 4.0 and 5.0, at which point the desired product is crystallized from the solution. The product is recovered by conventional methods, preferably by filtration, washed and dried to yield crystalline propylene glycolate with 1.0 to 1.6 moles of 1,2-propylene glycol bound per mole of cephalosporin zwitterion. . As shown below, the propylene of the present invention is a crystalline product, and is 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide] produced by an industrial method.
Impurities found in -3-(1,2,3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid are substantially absent.

Even when this is dispersed in water, its viability is not impaired, crystallinity and uniform dispersibility are not lost, and neither oiliness, lumpiness, sedimentation nor stickiness occurs. In the solid state, 5 is 0 even under normal severe test conditions, and the decline in viability is less than 10% even after storage for one month or more. The stability of this solid is much better than ethanolate at that temperature and even at 10,000 ℃. It is possible to remove excess propylene from the products of the invention by washing with acetone, methanol or ethyl acetate, but not with solvates such as acetone or ethyl acetate. In the examiner test, crystal 71 [D-Q-amino-Q-(
p-hydroxyphenyl)acetamide]-3-(1,
The efficacy of 2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid 1,2-propylene glycolate and subectrum is disclosed in a pending US patent (S.
No. 318,340) and the same level as that described in No. 318,340).

In some cases, the activity was slightly lower, but it is biologically inactive. . It was based on the inclusion of pyrene glycol. In the biological tests, the products were administered based on the effect of zwitterions, so the activities were at the same level within experimental error. In the treatment of bacterial diseases in humans, crystals 71 [D-Q-amino-Q-(p-hydroxyphenyl)acetamide]- can be administered orally or subcutaneously depending on the patient.
3-(1,2,3-triazol-5-ylthiomethyl)-3-cephemu 4-carboxylic acid 1,2-propylene glycolate was administered.

The dosage is about 5 to 200 o/k9/day, preferably about 5 to 2 o/k9/day and 3 to 200 o/k9/day.
The dose was divided into four doses. The dosage unit is e.g. 12525
0 or 500 o of the active ingredient diluted with a physiologically acceptable support, for oral administration in capsules or tablets containing solid ingredients, and also in aqueous dispersions for subcutaneous or oral administration. I mixed it. Preferred crystals of the present invention 7-[D-Q-amino-q-(p-hydroxyphenyl)acetamide]-3-(1,2,3 triazole-
The method for producing 5-ylthiomethyl)-3-cephemu 4-carboxylic acid monopropylene glycolate is as follows.

That is, (117-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-31(1,2.3-
Prepare an aqueous solution containing a water-droppable organic compound having a triazol-5-ylthiomethyl)-13-cephemu-4-carboxylic acid and a ketone functional group;
Prepare the H to about 4.5; dilute the solution with enough water to precipitate non-portable impurities; ■ separate the aqueous solution from insoluble impurities; ■ add enough water to the aqueous solution; This method consists of adding 1,2-propylene glycol to crystallize the desired monopropylene glycolate; and recovering the Tsukuda crystalline product.

Crystal 71 [D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid mono-propylene glycolate The most preferred method of the present invention for producing is as follows.

That is, (1'7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-
Triazol-5-ylthiomethyl)-3-cephem-
Prepare an aqueous solution (the pfl value of this aqueous solution is about 2.0 or less) consisting of a 4-carboxylic acid and a water-dropping keto acid selected from birvic acid or levulinic acid;
2' Adjust the pH of the solution to about 4.5; {3' Dilute the solution with a sufficient amount of water to precipitate the impurities; {4} Separate the aqueous solution from the impurities.し：■
A sufficient amount of 1,2-propylene glycol is added to the aqueous solution to crystallize the desired monopropylene glycolate, and the crystalline product is recovered.

Crystalline monopropylene glycolate was prepared by the method described above to obtain 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-131(1,2,3-triazole-
(5-ylthiomethyl)-3-cephem-4-carboxylic acid or a hydrate or solvate thereof, preferably a methanol solvate, in a sufficient amount of a hydrolyzable organic compound having a ketone functionality. to form an aqueous solution of cephalosporanic acid.

The ketone-containing water-lacquer organic compounds used are as follows. That is, keto acids such as acetone, pyruvic acid, levulinic acid, acetoacetic acid, ketone glutaric acid, salts of keto acids, and hydroxyketones such as dihydroxyacetone or fructose. Preferred compounds are water-bathable keto acids. The most commonly used compounds from the viewpoint of availability, price, etc. are pyruvic acid and levulinic acid. The starting material for cephalosporin may be any of the free forms, hydrates, or solvates of zwitterions, but methanolate is the preferred compound because it has the highest dissolution rate. If a complete solution cannot be obtained even after addition, the pH value is adjusted by adding an acid or a base to completely dissolve.

In a preferred method of practice, a water-soluble keto acid is used to lower the pH value of the aqueous reaction mixture to about 2.0 or less, thereby bringing the cephalosporanic acid into solution. If the use of keto acids alone does not result in a complete solution, key acids are added to increase solubility and the pH value is adjusted.
The aqueous solution is then converted to P by addition of a suitable acid or base.
Adjust the H value to approximately 4.5. When using the preferred keto acids, the pH of the acidic solution is raised to pH 4.5 by adding a base such as NaOH, preferably with vigorous stirring. The solution is then diluted with water to remove water-insoluble impurities.

Although the degree of dilution is not critical, it has been found empirically that a 1:1 dilution yields a product of high purity. Although the reaction temperatures in the above steps are not critical, these steps (especially the step of adjusting the pH value to 4.5 and the step of dilution) are preferably carried out at room temperature or lower.

The preferred temperature is about 5-20 oo to minimize the amount of inert impurities during dilution. After dilution, solid impurities are separated from the aqueous solution containing cephalosporanic acid and ketone group-containing compound by conventional methods such as filtration.

The exact form of the product in solution is not clear, but it is believed to be a loosely bound, soluble, physical complex of the cephalosporin zwitterion and the keto compound.
In any case, by using a keto compound, the PH4.
In Example No. 5, the cephalosporin zwitterion also exists as a solution, but insoluble impurities (containing substantially all of the coloring components) precipitate. After removal of solid impurities, the aqueous solution is preferably treated with activated carbon and then filtered before addition of propylene glycol. Add 1,2-propylene glycol in an amount sufficient to precipitate the monoglycolate,
The precipitated glycolate is then collected by filtration.

Then wash and dry. Monopropylene glycolate produced by the above method is substantially colorless, highly effective, and is a crystalline substance with excellent coloring stability and thermal stability.

Unlike other solvates, this compound does not lose its vitality, crystallinity, etc. even when dispersed in water, and does not cause oily formation, uneven dispersion, sedimentation, or stickiness, so it can be used as an aqueous dispersion. Particularly suitable for Considering that it contains inert propylene glycol, the monoglycolate of the present invention has been tested in both the examiner test and the in vivo test as described in the pending US patent specification (S. No. 318340). It has the same activity in both potency and spectrum as the other zwitterionic products. In the treatment of bacterial diseases in humans, crystalline monopropylene glycolate has been administered orally or subcutaneously, depending on the physician's choice.

The dosage was about 5 to 200 Ta'k9'My, preferably about 5 to 20 9/kg/ship, administered in 3 to 4 divided doses per day. Dosage units are, for example, 12 to 250 or 500 m9 of active ingredient diluted with a physiologically acceptable support, for oral administration in capsules or tablets containing solid ingredients, and for subcutaneous or oral administration in capsules or tablets containing solid ingredients. A hydration solution was also used. The present invention relates to crystal 71 [D-Q-
In addition to the method for producing amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid 1,2-propylene glycolate, the rational method described above 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]- which cannot be purified by conventional methods with a reasonable yield.
A method for purifying 3-(1,2,3-triazol-5-ylthiomethyl)-13-cephem-4-carboxylic acid is provided.

In the above method, 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-
(1,2,3-triazol-5-ylthiomethyl)-
When 3-cephem-4-carboxylic acid is used, cephalosporin is recovered in the form of pure crystalline 1,2-propylene glycolate.

The propylene glycolate is 1.0 to 1.6 mol, preferably 1.0 to 1.6 mol per mol of cephalosporin zwitterion.
Contains 0 moles of 1,2-propylene glycol. 1
, 2- If methanol is used instead of propylene glycol, a crystalline monomethanolate is obtained, which can also be used as a starting material for the preparation of crystalline propylene glycolate according to the invention.

The present invention also provides at least 9/skin of 30, preferably 125/skin
7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cefim-4-carboxylic acid 1,2- above the skin Propylene glycolate (in this propylene glycolate, cephalosporin zwitterion 1
1.0 to 1.6 mol per mole, preferably 1.0 mol of 1.2-propylene glycol) and its pH value is 2.8 to 5, preferably 2.8 to 5. 3
．． 5. To provide a stable aqueous dispersion effective in the treatment of bacterial diseases in humans.

The biological test is s. aureus20 squares (A.T.C.C.
．． 6538p) versus the standard 7-[D-Q-amino-Q-(p-hyde.

xyphenyl)acetamide]-3-(1,2,3triazol-5-ylthiomethyl)-3-cephemu 4
- using carboxylic acid 1,2-propylene glycolate,
I did it with great care. The potency of the sample was 82 kg/9 kg, and according to chemical analysis, 1,2-
Contains 16.7% propylene glycol and 0.3% water. Anhydrous zwitterion 7-[D-Q-aminoba-1 (p-
hydroxyphenyl)acetamide]-131 (1,2,
The titer of 3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid is 9 of 100 hc/h, containing 16.7% of 1,2-propylene glycol and no other impurities ( The sample (containing no water and excess or unbound propylene glycol) should have a calculated titer of 833 mc/9 ml. When the number of moles of propylene glycol bound per mole of zwitterion is 1.0 mole and 1.6 mole, the content of 1,2-propylene glycol is 14.1% and 20.8%, respectively. The molecular weight of the zwitterion is 462.38
It is. IR and N 8-position spectra were measured on the same sample.

The functional group data obtained from the spectrum is as follows. IR (KBr) 2400-3600 skin-1 (medium broad overlapping peaks) -amide, NH, NH30, OH1780 8-lactam C=0 1700 amide C=○ 1570 -COO- 1515 -aromatic C=C NMR (DMS Late DCI) 7.96ppm6 singlet IH. Hc6.7-7.
6 Multiplets. 4th, Hc5.7
Doublet, IH. Hc4.9-5.2 Multiprett, 2nd, Hd, He3.2-4.2 Multiprett, Chichi*, Hf, Hg, Hj, Hk1.1
BL-V64 from doublet, 4H*, Hn* integral value
The amount of propylene glycol per mole of zwitterion is 1.33 moles (18.3 wt% without moisture correction).

7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-131(1,2,3-triazol-5-ylthiomethyl)-13-cephemu-4-carboxylic acid synthesized according to the method of Example 5. The titer of monopropylene glycolate was calculated to be 9 of 865 mcta'.

Chemical analysis of the sample showed 15.6% 1,2-propylene glycol and 0.3% water. IR and NM
Previous measurements were performed on the same monopropylene glycolate sample and gave the following results.

IR (KBr) 2400-3600c -1 (medium broad overlapping peaks) -amide NH, NH30. OH1
780-13-lactam C=01705
-Amide C=○1570 -COO-1515 -Aromatic C=C
NMR (DMS○, rare DCI) 7.98 ppm 6 singlet, IH, Ha6.7-7.6 multiplet, 4 days, Hb5. Total Doublet, IH, Hc4.9-5.2 Multiplet, 2 days, Hd, He3.2-4.2 7 Triplet, 7 days, Hf, Hg, Hi, Hk1.1
Doublet, 3rd, BL-S from Hn integral value
The number of moles of propylene glycol per mole of 640 zwitterion was determined to be 1 mole.

The starting material D-(1)2-(p-hydroxyphenyl)glycyl chloride hydrochloride was synthesized in a reasonably high purity by the following method.

10.02 (approximately 0.06 mol) of D-(1)2-(p-
Hydroxyphenyl)glycine (US Pat. No. 348,975)
2) was slurried in 100% dioxane.

COC12 (phosgene) was passed through the slurry while keeping the temperature of the slurry at 50 to 0 while stirring. C
Passed through OC12 for 3.5 hours. A yellow solution was obtained.
The solution was purged with nitrogen to create excess COC12. 2. Add HCI gas to the solution. Insect time blew in. The solution was ignited and a small amount of the solution was diluted with Jenal to precipitate crystals, which were added to the batch as seed crystals. 2 of the solution
It was 0-2 and 0, and I was stuck at the foot for an hour. The obtained crystal D-(
1) A slurry of 2-(p-hydroxyphenyl)glycyl chloride hydrochloride was filtered to recover the product. The filter cake was washed with dioxane and methylene chloride and then dried over P24 in a vacuum dessicator. D-
(1) The yield of 2-(p-hydroxyphenyl)glycyl chloride hydrochloride was 7.3 days. IR-Good elemental analysis CI C Day N Theoretical value 31.93 43.14 4.09
6.37 Actual value 31.96 42.46 4.
22 6.56 Acid chloride analysis results Saddle chloride 9
8.6% Free COO Days None Free HCI None ○1Q-t-F.

8. Toxicarbonylamino-Q-(p-hydroxyphenyl)acetic acid was thoroughly mixed in a Trillo flask equipped with a reflux condenser, an overhead simulator, and a thermometer.
36 (0.05 mol) of D-(1)-p-hydroxyphenylglycine and 3.02 (0.075 mol)
A mixture of 120' of magnesium oxide and 120' of 50% aqueous dioxane was charged.

Stir the mixture for 1 hour and then 10.74 hours (0.74 hours).
075 mol) of t-butoxycarbonyl azide. The mixture is then boiled at 45-5000 ml while stirring.
Maintained under nitrogen for 7 hours. The solution was diluted 20 to 40 ml and extracted twice with 300 ml of ethyl acetate. water phase 1
Acidified to pH 4 with 0% citric acid solution and saturated with NaCI. The aqueous mixture was extracted three times with 400' of ethyl acetate. The solution was dried over Na2S04 and the solvent was evaporated. The residue was dispersed and purified using “Skell$olvsB” to obtain D-Q-t-butoxycarbonylamino-2-(p-
A solid of hydroxyphenyl)-acetic acid was obtained. Yield 10.
4 evenings (78.5%). 7-[D-Q-t-butoxycarbonylamine/-Q-(p-hydroxyphenyl)acetamide]-13-[1,2,3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid 7-amino-3 -(1,2,3-triazol-5-ylthiomethyl)-13-cephemu-4-carboxylic acid (6.0 evenings, 1
9. 8.5 M 1,1,1,3,3,3-hexamethyldisilazane (40.9 hmol) was added to a suspension consisting of 100 hmol of dry methylene chloride and 100 hmol of dry methylene chloride.

The mixture was heated to reflux for 4 hours to obtain a clear solution.
The solvent was evaporated and the residual oil was degassed under high vacuum at room temperature overnight. Dissolve the foamy residue in 85' dry THF and
After cooling to about -150 ℃, it was used for subsequent reactions. D-Q
-t-butoxycarbonylamine/-Q-(p-hydroxyphenyl)acetic acid (4.4 evening, 16.5 mmol) was added to 1
45' of dry THF.

The solution was tied up and cooled to -20°. N-Methylmorpholine (1.6 hours, lahmol) and isobutyl chloroformate (2.3 hours, 16.8 mmol) were added one after another slowly so that the temperature of the reaction mixture did not exceed about 11000 °C. The resulting mixture was then heated to -120 to -15
The lights were on for two ships at ℃. It was cooled to -200 and silylated 7-amino/-3-(1,2,3-triazole-
A THF solution of 5-ylthiomethyl)-3-cephem-4-carboxylic acid was added in one portion. The temperature rose to about -12 degrees. External cooling was stopped until the temperature rose to 0°C. When the temperature reached 0° C., the mixture was allowed to stand for 3 hours at 2-30°C using an ice machine.

External cooling was then stopped and the temperature increased to 200°C. Add 30 methanol and heat the female at room temperature for 15 methanol.
Lasted for a minute.

After evaporating the solvent under reduced pressure, the residue was suspended in 300' of ethyl acetate. The suspended solid was filtered to give 11.89. The ethyl acetate solvent was extracted three times with NaHC03 solution (5%). The hard carbonated extracts were combined, phase separated with ethyl acetate, and adjusted to pH 2 at 42.5% day 3P03.
．． Acidified to 5. The phases were shaken and then separated. The ethyl acetate solution was then dried by passing over Na2SO4 and then evaporated to approximately 15-20'. This solution was then added dropwise into cyclohexane (~40 kg) in an Erlenmeyer flask. 1/Kitsuneterama After heating, the precipitated solids were collected in the furnace.

The recovered 7-[D-Q-t-butoxycarbonylamino-Q-(p-hydroxyphenyl)acetamide]-131(1,2,3-triazol-5-ylthiomethyl)-
The 3-cephyl 4-carboxylic acid was air dried. Yield 1
．． 76 evening. 7-[D-Q-t-butoxycarponylamino-Q(p-hydroxyphenyl)acetamide]-3
-(1,2,3-triazol-5-ylthiomethyl)
-3-cephem-4-carboxylic acid 3.5% purity 98~
Dissolved in 100% HCOOH80 magnetic and stirred it at room temperature for 2 hours.
I sat down for a while.

The HCOOH was evaporated under reduced pressure (using an aspirator, temperature below 40 pm) and finally azeotropically distilled three times with 30 ml of toluene. The solid was dried over P205 under high vacuum overnight. A foam of 3.5 hours was obtained.

The foam was quenched with 300 parts water and methanol (8:2). Heat the solution in an oven (solid content 0.3 hours), and heat it for 7 hours.
Treated with 00mp activated carbon "DarkoKB", filtered through diatomizer ("Celite") and freeze-dried to give crude 7-[D-Q-amino-Q-(p-hydroxyphenyl) Acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-yluponic acid was obtained. 0.2 99% of the above crude product with 6
It was dispersed in methanol and heated to boiling in a test tube.

Heating was immediately stopped and seeds were added to the melt. The melt solidified into crystalline form. Thus, from 0.400 μm of crude material to 0.211 μm of 71[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-
Triazol-5-ylthiomethyl)-3-cephem-
4-carboxylic acid was obtained. Product 5/0.1 skin, P20
The two legs were dried on 5. m. p. 72000 decomposition. I
R and NMR spectra were consistent with the structure. NMR
It was found that 1/3 mole of methanol was present. Elemental analysis/calculated value (C, 8th, 8N605S2/day 2
0.1/3CH30H):C,44.83:Sun,4.3
8; N, 17.10: S, 13.09 Actual value: C, 43
．． 97: Sun 4.36; N, 15.84: S, 6.18.

6.5 evening 7-[D-Q-t-butoxycarbonylamino-Q-(p-hydroxyphenyl)acetamide]-131(1,2,3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid was dissolved in ~100% formic acid 176 under anhydrous conditions.

The mixture was allowed to stand at room temperature for 2.5 hours. It was evaporated under reduced pressure to remove 125 skins. The residue was an amber oil. The oil was then co-precipitated with 70% of triene under reduced pressure three times. The residue was dispersed in 700 g of water-methanol (80:20) and stirred until most of the solids were dissolved. It was then filtered. The furnace liquor was treated with 15 days of "Darko" activated carbon. Activated carbon
The solution was then lyophilized and its weight was 2.415 kg. It was recrystallized in the above method with 0.200 kg of
7-[D-Q-amino-(p-hydroxyphenyl)acetamide)-13-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid was obtained. NMR showed the same structure as the desired product. Also, i'3 moles of methanol were bound. Elemental analysis/calculated value (C.8 days, 8N605S2, days 20, 1 no 3 CQOH): C, 44.83: days, 4.3
8; N, 17.10; S, 13.09 Actual value: C, 45
．． 774436; Sun, 444,434:N16.61;
16.52; S, 13.01, 13.017-amino-
Acylation of BL-S640 of 3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid (7-TACA) was performed using D-(
1) -p-Hydroxyphenylglycyl chloride hydrochloride is used.

The yield of BL-S640 methanolate was about 45% based on biological titer. 15% remains in the mother liquor; 25%
is an insoluble solid of unreacted 7-TACA, and also contains decomposed products such as decomposed B-lactam.

The method requires silylation of 7-TACA with HMDS, followed by acylation with acid chloride hydrochloride at 0-5°C and quenching with methanol.

Furthermore, methylene chloride is removed from the reaction mixture, and the methanol solution is treated with activated carbon. The furnace solution is concentrated under reduced pressure, then adjusted to pH 4.8-5.0 with concentrated aqueous ammonia, and seed crystals are introduced for crystallization. Reaction formula 313.3 7-TACA (U.S. Patent 3,759,904) 2224
62.45 Reaction operation 1 1000 units (3.20 mol) of 71 TACA is added to 25
Add to the dry methylene chloride (KF. day 20 <0.01%).

The generated slurry was tied to a rope of 1245' (approximately 5.3
Add 5 moles) of HMDS to the slurry. 2 Heat the slurry to reflux while blowing dry nitrogen gas into it.

Refluxing is continued until no undissolved material is observed and the solution is completely dissolved. Reflux continues for 12 to 2 minutes to obtain a concentrated solution. 3
After the silylation is complete, the solution is cooled to about 15-20qO, and 320' of DMA/ACI (30% in methylene chloride) is added.
%), then DMA (dimethylani1') 48
Add 21.8 tbsp and imidazole.

The reaction mixture is cooled to 0-5°C and 797 (3.60 mol) of D-(1)-p-hydroxyphenylglycyl chloride hydrochloride is added in 5 portions over 1 hour. The slurry is heated at 0-5° C. for 10-12 hours to dissolve the acid chloride. 4 The reaction mixture was heated to 2000 ml over 3 hours.
Heat gradually to 2000 and hold for 2 hours.

Care must be taken to ensure that the acid chloride is completely dissolved. 5 8.3 The dry methanol (KF, day 20<0.
01%) to the solution within 1 minute, stirring well.

The mixture is stirred for 10 to 15 minutes and then quickly heated in a furnace to remove unmelted substances (for experimental materials, filtration is carried out in a Buchner funnel and the cake consists of 2 parts methylene chloride and 1 part methanol). (Clean with a cleaning solution.) It is necessary to perform the furnace cleaning quickly, and for this purpose, it is necessary to assemble the furnace filtration device in advance. After passing through the furnace, solids precipitate from the furnace liquid and washing liquid. It is not clear whether these solids are product (probably hydrochloride). During this filtration, the separated solids contain some product 7-TACA and decomposed 7-TA.
Contains CA. The cleaning solution is methanol-methylene chloride (
1:2), and it takes about 10 minutes. 6 This furnace liquor is concentrated under reduced pressure to remove methylene chloride, and then the required amount of dry methanol is added.

The solution was concentrated to 15-18 ml and 600 ml "Darko"
Add ``KB''.Light the lamp for 20-25 minutes, then add the slurry to ``Dicaljt''.
〆) Filter with pre-coat.

Then wash the cake thoroughly with 8.0 methanol.
This operation usually yields a yellowish-colored furnace liquid. 7 1 part furnace liquid
Concentrate under reduced pressure from 2.0 to 13.0 pm and add 480 ml of deionized water.

The pH should be between 2.4 and 3.2. Concentrated ammonia water was gradually added dropwise to the solution over 30 minutes until the pH was adjusted to 4.
8 to 5.0. If you expand the scale of the laboratory, it will be 42.
0-440' of ammonium hydroxide should be required. Seed the solution and adjust the pH value to 4.0. The pH was adjusted to 2000, and then the slurry was
The mixture was roasted for 1 hour at 0°C and further cooled for 1 hour at 0°C. In the laboratory, after incubating in an ice bath for 3 hours, the beaker is placed in ice and stored overnight in a freezer. - Crystal growth was always observed on the sides of the beaker after standing overnight.

At this point, it was not clear how much time the solution should be allowed to stand, but analysis of the remaining liquid revealed that leaving it for about 3 hours was insufficient. The precipitated product was collected by filtration, washed with the same amount of methanol as the product, and dried at 45°C. Typical yield is as methanol solvate of 7-[D-Q-amino-Q-(p-hydro.xyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid. It is 750-770 evening. Anhydrous conditions must always be maintained during this reaction operation.

When water is mixed in, the silyl ester is hydrolyzed, making it difficult to proceed with subsequent acylation. The present invention will be explained below with reference to Examples, but the present invention is not limited thereto.

All temperatures are in °C. “Tween8 people”P
It is a mixed polyoxyethylene ether mixture of partial oleic acid ester of anhydrous sorbitol, also known as oleic acid. The 1,2-propylene glycol used was Propylene Glycol U. S. P. Tetrahydrofuran is abbreviated as THF. "Skell $olve B" is a crab-based petroleum ether with a boiling point of 60 to 68 oo, and its main component is n-hexane. Example 1 Production of crystal 71 [D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-31-(1,2,3-triazol-5-ylthiomethyl)-3-cephemu 4-carboxylic acid metalate 1 50 evenings 7-[D-Q-amino-(p-hydroxyphenyl)acetamide]-
3-(1,2,3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid 95 above 250
% V/V methanol/water (95% methanol) solution at 22-25 CC.

2 While lighting the solution, quickly add concentrated hydrochloric acid to bring the pH to 1.3.
~1.5.

A solution or simulated solution is obtained. 3 PHI with triethylamine. Set it to 7.

4 Add activated carbon (Daric G-60'') for 7.5 hours and slurry for 0.5 hours.

5. Heat the activated carbon in a furnace and wash it with 75 minutes of methanol.

The cleaning fluid is integrated with the furnace fluid. Steps 2, 3, and 4 must be completed within 5 hours. 6. Immediately ignite the combined furnace fluid and cleaning fluid. Add triethylamine over 5 minutes to reach pH 4.5. Crystallization begins in 1-3 minutes.

Slurry the mixture for 1 hour. 7. The crystals are collected by filtration, washed with 100M methanol, and vacuum dried at 56q0 for 24 hours.

Biochemical yield 75-90%: Biochemical titer 850-90 hcg/o; NMR-IR: Consistent with conjugate of 1 mole methanol: % average OKF = 2-4.0 crystal BL-S64
01,2-Propylene glycolate production 1 25-7-[○-1Q-amino-Q-(p-hydroxyphenyl)acetamide]131(1,2,3triazol-5-ylthiomethyl)-3-cephemu 4 - Carboxylic acid methanolate (prepared by the above method) at 150-200%
20 to 75% V/V propylene glycol-aqueous solution of
Slurry at 25°C.

2 Adjust the pH value to 1-1.2 with concentrated hydrochloric acid to obtain a pseudo solution.

3. Gradually add triethylamine (TEA) to pH.
7 to 1.8. Add Darco G-6 and slurry the mixture for 0.4 hours.

Remove the activated carbon in a furnace. It is desirable to carry out this furnace gradually. 576 oven paper is preferred.
The activated carbon filter cake is washed with 40 g of 75% V/V propylene glycol-water temperature mixture. The wash solution is integrated with the solution. Steps 2, 3, and 4 must be completed within 5 hours. 5. Add triethylamine to the combined furnace solution and cleaning solution obtained in step 4 over 1 minute while lighting the mixture to adjust the pH to 4.5.

Crystallization begins in 1-3 minutes.

Slurry the mixture for 1 hour. 6 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]
-3-(1,2,3 triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid propylene glycolate is recovered by filtration.

Filtration should be done slowly and SS No. 125-15.0c. It is recommended to use 576 furnace paper. 50 crystals
The wings were washed sequentially with 75% propylene glycol, 50% methanol, 50% acetone, and vacuum dried at 56°C for 24 hours. Productive yield 80-95%. 7-
Properties of [D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid propylene glycolate a Biological potency = 750-7
9 ratio hcg/gu. b m-NMR = consistent with the structure of the target product with 1.3-1.5 mol of propylene glycol bound (propylene glycol content 17-19%).

No loss of 3-triazole side chain is observed.

c % moisture, KF = 1-3.0 d Crystallinity = 100% Triangular, microcrystalline e M. P=
182-1840 (hot plate) f [Q] Ginseng (C = 1%; INHCI) = +530g
Water dropability = 2300 and 9' secretion/ratio ○h of about 10 Viability loss rate at high temperature: 100°C, 24 hours = <6%; 4 glue time = <12%, 1 month = <10% Example 21. Charge 1.5 g of propylene glycol and 1.5 g of deionized water to a suitable container equipped with a pot and pH control device.

2 To the above propylene glycol-water (1:1) mixture was added 1000 g of 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3 triazol-5-ylthiomethyl). -3-Cephem-
Add 4-carboxylic acid methanolate.

3 While thoroughly lighting the above slurry, mix 15 qo with 25 qo.
By adding Iso HCI of 425 people over a period of time, the multiplication value is 0.
．． 9 to 1.3.

A tonally clear solution should be obtained. 4 Immediately T
Adjust the pH value to 1.4-1.7 with EA (triethylamine).

It only takes 20 to 30 people. A small amount of white solid precipitates out. This precipitate is considered to be p-hydroxyphenylglycine or a derivative thereof. 5. Treat the solution with "DarCOKB' for 50 days.

Bake the slurry at 2500 for 15 minutes. 6. Remove the activated carbon with a pre-coated "Dicalite" filter.

The furnace area requires 1.3 mm/day, and on a laboratory scale it is desirable to apply a vacuum. It is desirable that the furnace overspeed be slow, so that the surface of the cake is finally scraped. Furthermore, pressure furnace filtration is also expected to give favorable results.
The activated carbon cake is washed with 1400 g of a 7:3 propylene glycol/water solution. Store this washing solution separately. 7 Pass the furnace liquid through a suitable sterile filter and place it in a suitable sterile container.

The furnace area is 1.3/h. Wash the filter pad with the wash solution from step 6 and then again with 1000 M sterile propylene glycol-water mixture (7:3). 8. Add 1.75 ml of sterile propylene glycol to the sterile container from step 7.

9. Gradually adjust the pH value of the solution obtained in step 8 to 4.1-4.3 over 20-30 minutes with 3000ml sterile TEA under vigorous lighting.

10 Light the slurry continuously at 25o0 for 4-5 hours.

The slurry is stable even when stored overnight. 11 Filter the sterile crystals and wash the cake with 1000' of sterile propylene glycol/water (7:3) and 1000' of sterile ethyl acetate.

12. Slurry the sterile crystals again in 2000 g of sterile ethyl acetate to remove excess propylene glycol.

13 Collect the solid by filtration and wash the cake with 1.2 sterile ethyl acetate/methylene chloride mixture (1:2).

14 Dry the product in a 50q0 vacuum dryer for one batch.

The yield of crystal 71 [D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid 1,2-propylene glycol is 820-9
It is the 10th evening.

15 Product analysis: Propylene glycol: 7-[D
1-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephemu 1.2 to 1.3 mol per mol of 4-carboxylic acid (NM depending on the town).

Pyridine: 0.04% or less (according to VPC). Biological titer: 800-85 heg/9. Note 1 Raw material BL-S
640 methanolate contains 0.4-0.6% pyridine.

2. The white precipitate generated in step 4 is
The filter can be preheated with a sintered glass filter precoated with ``Dicalite''.

Doing this will make it easier to filter the activated carbon. 3 7th color
-[D-Q-amino-Q-(p-hydroxyphenyl)
Acetamide]-3-(1,2,3 triazole-5
-ylthiomethyl)-3-cephemu-4-carboxylic acid methanolate requires treatment with a larger amount of activated carbon.

This makes filtering more difficult. 4 Steps 6 and 7 need to be completed as quickly as possible.

Sterile furnace fluid should not be stored for more than 5 hours. If necessary, a portion of sterile TEA can be added to the furnace fluid before cleaning. 5. The sterile propylene glycol in step 8 is sterile; it is desirable to sterilize it by heating it at 8000C for 30 minutes after passing through the door.

and sterile 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-
Cool to 25° C. before adding to the carboxylic acid solution. Alternatively, sterile propylene glycol may be substituted with the sterile oven solution of step 7. Example 3 Sterile crystalline subcutaneous grade 7-[D-Q-amino-Q-(p
-hydroxyphenyl)acetamide]-3-(1,2
, 3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid propylene glycolate 1
7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephemu-4-carboxylic acid methanolate or recrystallized 7- [D-
Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3triazol-5-ylthiomethyl)-13-cephem-4-carboxylic acid propylene glycolate in 20-25q○ of 150-200 Slurry in 75% V/V propylene glycol-water solution.

2 Add concentrated hydrochloric acid to adjust the pH to ~1.2 to prepare a solution or pseudo-solution.

3. Gradually add triethylamine (TEA) under vigorous lighting to adjust the pH of the solution to 1.6-1.8.

Add 4-5 ounces of Darco G and bring the mixture to 0.
Make it into a slurry for a while.

Activated carbon is removed by filtration. The week of the furnace is late, SS of 18.5 skin
No. 57 mochido paper is preferred. Wash the activated carbon filter cake with 40 ml of 75% V/V propylene glycol/water solution. Add cleaning solution to furnace fluid. 5. Pass the combined furnace fluid and cleaning fluid through a 0.22 micron string hole filter and store in a sterile container or tank in a sterile area.

6. Add sterile triethylamine to the sterile solution from step 5 over 10 minutes with vigorous stirring until the pH value is 4.5.
shall be.

Crystals form in 1 to 3 minutes.

The mixture is slurried for 1 hour. 7. Obtain sterile crystals in a sterile oven.

Mito Uzu is slow (12.5-15.0 skin, SS No. 57
6 oven paper is preferable). The crystals are washed with 50 μm of sterile 75% propylene glycol, 50 μm of sterile methanol, 50 μm of sterile acetone, and vacuum dried at 56 μm. Biochemical yield 80-95%. 8 Sterilize the sterile crystals and grind them to 200 mesh.

Properties of 7-[D-Q-amino-Q-(p-hydroxyphenyl)-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid propylene glycolate a. Biological Titer = 800-86 skin c/
9. b IR-NMR = consistent with the target compound containing 1.3 to 1.5 moles of propylene glycol.

Propylene glycol content 17-19%. No loss of 3-triazole side chain is observed.

c Crystallinity = 100%, triangular microcrystals d Moisture (%), K. F. =1~3.0eM. P. 182-184oo (hot plate) f [Q] Customer (C=1%; IN-HCI) = 1530g Water solubility = about 9 of 10/wing -23qoh Thermal stability: 100qo, 2
4 hours = <6% loss; 100°C, 4 songs time = <12% loss; 56q○, 1 month = 10% loss.

Example 4 Production of purified BL-S640 monomethanolate 1 10
Pour 0.0 ml of BL-S640 methanolate or 1,2-propylene glycolate into 300 ml of water and quickly light up.

2 Add 80 ml of levulinic acid.

3 Shake the solution vigorously and gradually add concentrated hydrochloric acid to adjust the pH.
The value is set to 0.8 to 1.2 to obtain a solution or a pseudo solution.

Cool the solution to 20-25 qo. 4 While vigorously stirring the solution from step 3, add 40% sodium hydroxide to bring the pH to 4.5.

Denden is generated once at pH 2.0 to 3.0, and then the pH
4-4.5 to dissolve again. The temperature must not rise above 27°C. 5 Cool the solution to 4-10%,
Further, stir the solution very vigorously and add 4 to 1,000 g of water to make 500 g.

Precipitate is formed. 6 Heat the mixture at 4-100° for 5 minutes.

The precipitate (x) containing colored components and impurities (including the detriazolized product of BL-S640) is removed by filtration. Wash the sediment with 50ml of water (this washing liquid must not be mixed with the furnace liquid). Then wash with 75' of methanol (also do not mix with furnace liquor). Vacuum dry at 5℃ for 2 mornings.

A pale brown solid (0-500 units/female) of 5-15 hours is obtained.

7 Add Darco G60 or KB activated carbon from 15th day to the furnace liquid (Step 6) of Tanden (X).

The mixture was heated at room temperature to 0. Asa Terama Kenhan. 8. Remove activated carbon by filtration and wash with 40' of water.

Add the wash water to the furnace fluid. 9. The furnace liquid is sterilized through a filter with a pore size of 0.22 microns.

Steps 4-9 should be completed within 4 hours. 10
Sterile, equal volume (approximately 1.5 liters). Add pyrogen-free methanol to the pH 4.5 solution obtained in step 9 with gentle lighting.

Crystals form in 1 minute.

Maintain pH at 4.5. 11 Heat the mixture to 18-23o
I pretended to be at o for 1 hour. 12 Filter off the shiny white crystals and add 175' of sterile 50% methanol, 3
Wash with sterile methanol at 56 qo and vacuum dry for 2 hours at 56 qo.

Yield 65-75 min (biological yield 70-80%). Example 5 Production of BL-S640 monopropylene glycolate 1 100 ml of BL-S640 methanolate or 1,2-propylene glycolate is charged into 300 ml of water and vigorously vortexed.

2 Add 8M levlinisoic acid.

3 While vigorously stirring the chicks, gradually add concentrated hydrochloric acid until the pH value reaches 0.
8-1.2 to obtain a solution or simulated solution.

Cool the solution to 20-25q0 if desired. 4 While vigorously stirring the solution obtained in step 3, add 40% sodium hydroxide over 5 minutes to adjust the pH value to 4.5 (pH
At a pH of 2.0 to 3.0, makuraden is formed, and then at a pH of 4 to 4.
5).

The temperature should not rise above 270 degrees. 5 Cool the solution to 4-1oo0 and add 4-10q while stirring vigorously.
Add o of water to make 500 [.

Next, electricity is generated. 6. Stir the mixture at 4-1000 for 5 minutes.

Coloring components and impurities (Detriazo W of BL-S640
The next starch (x) containing chlorides) is removed by filtration. The precipitate is washed with 500ml of water (this wash must not be mixed with the furnace liquor), then with 75' of methanol (also not mixed with the furnace liquor), and then washed with 500ml of methanol (also not mixed with the furnace liquor).
Vacuum dry for two periods at 0. A light brown solid (0 to 50 units/female) of 5 to 15 days is obtained.

7 15 evening Da [coG60 or KB sinking (X
) to the furnace solution (step 6) and bring the mixture to room temperature.

Lights are lit in the morning. 8. 40 Z
Wash with water.

Wash water is integrated with furnace fluid. 9. The furnace solution is sterile filtered through a filter with a pore size of 0.22 microns.

Steps 4-9 should be completed within 4 hours. 10
Add an equal volume (approximately 1 night) of sterile, pyrogen-free methanol to the pH 4.5 solution obtained in step 9 with gentle stirring.

Crystals form in approximately 1 minute. Maintain fences at 4.5.
11 Stir the mixture for 1 hour at 18-2°C.

12 The shiny white crystals were removed by filtration, and 175'
Wash with sterile 50% methanol, 300 °C of sterile methanol, and vacuum dry at 56 °C for 2 hours.

13 Yield 70-80% (biological yield 75-85%/
BLS640 Monopropylene Glycolate).

The monopropylene glycolate sample obtained in Example 5 was analyzed and the following results were obtained. a Biological titer = 865mc
g/ob IR-NMR = consistent with the structure of the target product in which 1 mole of propylene glycol is bound per mole of cephalosporin zwitterion c % water, KF = 0.3 d Crystallinity = good, rod-shaped crystals e [Q ] Customer (C = 1%; INHCI) = 15590f
Propylene glycol % by chemical analysis = 15.6g
Other solvents = 0.1% h UV absorption spectra (in 0.1NHCI): m
ax=227 nm (o=284) and 27 m (Q=
16.6) A monopropylene glycolate sample was analyzed by powder X-ray diffraction according to the following method.

The resulting sample was highly crystalline, giving 39 measurable diffraction lines.

The relative strength between the surfaces is as follows. Details of the X-ray diffraction measurement method are as follows. That is, a small amount of the sample was sealed in a low dispersion glass capillary tube with a diameter of 0.2 and inserted into a Debyshiler powder diffraction camera with a diameter of 114.6.

Exposure time is NorelcoX operated with 3 songs V-2 dishes A
It takes 4 hours with a line generator. A standard focus steel target X-ray tube (CMQ, wavelength input - 1.5418A) was also used. Using Kodak No-Screen X-ray film and Kodak liquid X-ray developer at 20°C. Developed for 3 minutes.

A certain amount of sodium fluoride was added to the sample to serve as an internal standard.

Pure NaF was also measured in the same manner. The film is Norel
The position of the diffraction ring was read with an accuracy of 0.005 using a codevice error film reader. The data was corrected for film shrinkage, and the interplanar spacing was calculated. A computer was used for the calculations. The accuracy of the obtained d distance is 1%
Within Strength is Joyce-LoebleMark
It was measured using an mc microdensitometer.

(Scan ratio 5:1, 0.1o.D.wed bag). Relative intensity was expressed on a scale of 1-100. Example 6 Production of purified BL-S640 monomethanolate 1 10
Dissolve 0.0 ml of pyruvic acid in 250-300 ml of water. Add 2.100 ml of BL-S640 propylene glycolate or BL-S640 methanolate to the above solution over 5 minutes with vigorous stirring.

A solution or pseudo solution with a pl of 12.0 is obtained. 3
Cool the solution to 1000 ml.

4. Add 40% sodium hydroxide over 5 minutes to the solution obtained in step 3 with very vigorous loss, p.
The H value is set to 4.5.

At pH 2.0 to 3.0, Makuraden is generated for a day, then at pH 4.
Dissolve again at ~4.5. The temperature must not rise above 270°C. 5. Cool the solution or approximate solution to 8-1 to 0, and add water cooled to 8-1 to ○.

Pillowden precipitates. 6. Boil the mixture for 3 minutes at 8 to 1 ○. Makuraden (Iso) (This is a coloring ingredient and BL-
(containing impurities including detriazolated products of S640) are removed by vacuum filtration. Furnace velocity is small. Place the oven paper and cake in 300 ml of methanol,
Hold time. Air dry the oven paper and cake, scrape off the solids, and vacuum dry at 50 qo for 2 hours. 5-2
Light brown solid (titer 200-70 units/female) after 5 days.

Save the cake. 7 DarcoG60 on the 15th evening
Alternatively, add KB to the furnace solution in step 6.

The mixture was heated at room temperature to 0. Insect time lights up. 8. Remove activated carbon by filtration and wash with 40' of water.

Add the wash water to the furnace fluid. 9. Filter the furnace solution aseptically through a micropore filter with a pore size of 0.22 microns.

Steps 4-9 must be completed within 4 hours. The temperature must be maintained below 24 qo in steps 7-9. 10 Add an equal volume (1 drop) of sterile, pyrogen-free methanol to the solution obtained in step 9 at pH 4.5, while gently stirring.

Crystals are formed for about 1 minute. Maintain pH at 4.5. 11 Boil the mixture at 10-20 oo for 1 hour.

12 The shiny white crystals were removed by filtration, and 175'
Wash with sterile 50% methanol at 450 °C and sterile methanol at 450 °C and vacuum dry at 5600 °C for 2 hours.

13 Yield 50-60 days [Biological yield: BL-S640
Monomethanolate (biological titer 930-960 units/
60-75% of yield9)].

Example 7 Preparation of BL-S640 Mono-Propylene Glycolate 1 Dissolve 100ml of pyruvic acid in 250-300ml of water.

2. Add 100 mg of BL-S640 propylene glycolate or BL-S640 methanolate to the above solution while stirring vigorously for 5 minutes.

A solution or pseudo-solution with a pH of 2.0 is obtained. 3 Cool the solution to 10 qo.

4 Add 40% sodium hydroxide to the solution obtained in step 3 while stirring very vigorously to bring the pH value to 4.5.
(At pH 2.0 to 3.0, starch forms once, and then dissolves again at pH 4 to 4.5.)

At this time, the temperature must not be raised above 270°C. 5
Cool the solution or simulated solution to 8 to 12 degrees Celsius, add 8 to 1 year old water while stirring it vigorously, and heat to 500 to 60 °C.
0 kites

Precipitate is formed. 6. Soak the mixture at 8-120 for 3 minutes.

The starch (which contains impurities including color components and detriazolates of BL-S640) is removed by vacuum filtration. Furnace velocity is small. Aspirate the filter cake until dry. 30 hearth paper and cake
0' methanol and held for 4 hours. Air dry the furnace paper and cake, scrape off the solids, and reduce to 50 qo.
Vacuum dry for 24 hours. Light brown solid (5 to 25 minutes)
A titer of 200 to 70 mountain units/9) is obtained.

Save the cake. 7 DarcoG60 on the 15th evening
Alternatively, add KB to the furnace solution in step 6.

The mixture was heated at room temperature to 0. Insect time boundary hansuru. 8. Remove the activated carbon by filtration and wash with 40 ml of water.

Add the cleaning solution to the furnace fluid. 9. Aseptically filter the furnace liquid through a micropore filter with a pore size of 0.22 microns.

Steps 4-9 must be completed within 4 hours. The temperature must be kept below 2400 in steps 7-9. 10 Add an equal volume (approximately 1 night) of sterile, pyrogen-free propylene glycol to the pH 4.5 solution obtained in step 9 under gentle lighting.

Crystals precipitate for about 1 minute. Maintain pH at 4.5. 11 Boil the mixture in 10 to 20 circles for 1 hour.

12. Remove the shiny white crystals during the first week of illness.

Add it to 175' sterile 50% propylene glycol
The sample was washed with aqueous solution, further washed with sterile Meknol of 450 g, and vacuum dried at 56 C for 24 days. 13
Yield 55-60 min (60-70% biological yield based on crystalline BL-S640 mono-propylene glycolate).

Example 8 Intramuscular use 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-131(1,2,3-triazol-5-ylthiomethyl)-13-cephem-4-carboxylic acid ( Hereinafter referred to as BL-S 0) propylene glycolate (micronized) drug [Label indication is 250 o/secretion BL-S64 Erosion (as BL-S 0 glycolate)] *
Labeling is 250 9/Imitation BL-S640 activity (as propylene glycolate).

The required amount of BL-S640 propylene glycolate is calculated as follows. This amount needs to be increased by each of the following factors.

1 Add a larger amount in preparation for the storage period (stability) 2
Syringe and needle holdship 3 Equipment variations.

BL-S640 propylene glycolate (micronized) for intramuscular use [labeled as 250/9/desk BL-S640 (
As BL-S640 propylene glycolate)]1
The BL-S640 propylene glycolate used must be sterile and pyrogen-free.

Additionally, it must be handled aseptically throughout the process. 2 BL-S640 propylene glycolate is micronized in a sterile micromachine. 3. Add the mixture of sterile microscopic L-S640 propylene glycolate and sterile table salt to a sterile PaMrsonKellyV.
Charge to a blender (equipped with a bar for fluid addition).

It is necessary to sterilize the blender by spraying it with peracid and ethylene oxide before use. Tazushi blender
Care should be taken to avoid condensation of gas within the tank. Condensation is achieved by maintaining appropriate pressure and temperature. Run the blender on intensification for 30 minutes to ensure sufficient initial mixing. 4 Lecithin, Methyl and Propylparaben, Tween80 and Povid
One is dissolved in methylene chloride of 1/5 weight of BL-S Mouse 0.

5. Sterilize the solution from step 4 under pressure under aseptic conditions through a sterile 0.22 micron pore size microfilter and store in a suitable sterile container for sterile storage.

6. Add the required amount of the sterile, pyrogen-free methylene chloride solution prepared in step 5 in 5 portions using the "Liquid Addition Tool" of the blender.

Maximum lighting is performed using the intensifier bar for 2 minutes after each addition, followed by normal blending for an additional 13 minutes.
At the end of each addition, remove the pressure applied during blending (
(monitored with a pressure gauge attached to the outside wall of the blender), reduce the pressure and remove the methylene chloride. This operation must be repeated to completely remove the vapors. To improve the efficiency of evaporation and steam removal, hot water may be circulated around the outside wall of the blender to raise the salt temperature to 1150F. 7
After all the solution has been added and the steam removed appropriately, the mixture is dumped from the blender and transferred to a tray for drying.

Cover the mixture, place it in a marked tray, and place it in a hot air oven to dry for 6 hours. The temperature of the heated air is 130
It should be 0F or higher. After heating for 6 hours, heating is stopped and air is circulated over the tray to completely dry it.

8. Pulverize the coated product using the equipment used in step 2 to satisfy the following conditions.

200 mesh screen residual rate max. 0.1%9 Collect product in sterile container container.

10 Fill an appropriate amount of coated BL-S640 propylene glycolate into an officially certified silicone lubricant bottle under sterile conditions.

Example 9 Preparation of BL-S640 Propylene Glycolate Capsules (BL-S64 Erosion per Capsule is 9 of 250) BL-S640 Propylene Glycolate + 0.3148 U.L. Lactose per Capsule. S. P.O. 0
087 Magnesium tastearate U. S. P.O. 00
Total amount per capsule: 0.3250mg
- S640 propylene glycolate is 262.5%, which is 5% excess compared to the indicated amount of 250%.

Calculation of BL-S640 propylene glycolate activity is performed using the following formula. 0.2625×1000=〇. 3,4
834 (Amount per capsule) Note: BL-S640 propylene glycolate needs to be finely divided to pass through a 60 mesh screen before being blended with other ingredients and filled into capsules.

Example 10 1. M. 3 formulations of BL-S640 propylene glycolate 300 for dispersion BL-S640 propylene glycolate 100.3 per ampoule
59 Tween 80 (sterile, micronized) Sodium chloride (sterile, micronized) 0.0024 Tween80
0.0012 Talecithin
0.0024 Tapolypinylpyrrolidone (Povidone) Total amount per ampoule of 0.0060 to 10 0.3710 to 10 This amount corresponds to 9 out of 300 of the activity of BL-S640 9.
When adding 7' of water, 9' of BL-S640.30
A dispersion having an activity of .

Example 11 1. M. Dispersion BL-S640 Propylene Glycolate Preparation for 1.0 T BL-S640 Propylene Glycolate + 1.196 T (sterile, micronized) Sodium chloride (sterile, micronized) 0.008 T per ampoule
ween80 0.
004 Talecithin 0.
008 Polyvinylpyrrolidone (Povidone) 0.020
(total of above) (1.236 evenings) Additional amount of above composition for VNS holdup + 10
Total amount for ampoule of 0.349 5 1. Madara 5 Yu +
This amount is 10 V, which corresponds to 1.0 BL-S
NS means ampoule, needle and syringe.

Adding 3.7' of water for injection gives a suspension of 9' of BL-S640 activity 250.

Regenerated 250 female/'active BL- produced as described above.
S I 0 Propylene Glycolate 1. M. Storage stability was determined for the suspension.

The results are as follows. The dry powder was stable for at least 4 months at 5 and 0.

The above suspensions are improved by the addition of small amounts of non-toxic, pharmaceutically acceptable polycarboxylic acids, such as citric acid.

The amount of acid used (acid, of course, is added in dry form to the mixture of other solid components) is determined by the pH of the dispersion when dispersed in the liquid.
The amount is sufficient for the H value to be in the range of 2.8 to 3.5.
The pH value of the dispersion without addition is 4.4-5.
The improvements made by adding acid are the color tone, that is, the suppression of an increase in undesirable coloring components due to standing, and the suppression of deterioration in life quality due to standing. Effect of oral administration in beagle dogs Three beagle dogs (average weight: 8.2 kg, 0.4 k9) were given 200 mesh 71 [D-Q-amino-o-(p-hydroxyphenyl)acetamide]-3.
-(1,2,3triazol-5-ylthiomethyl-
3-cephem-4-carboxylic acid propylene glycolate (vitality 82 skin/k9) was orally administered in a hard gelatin capsule with a 30 female activity/k9.

The results shown in Table 1 are 30 o/X9 71 [D-Q-ami/-Q-(p-hydroxyphenyl)acetamide]
This is the average concentration in the blood plasma of a beagle-sized dog to which 1-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid propylene glycolate was orally administered. Approximately 40% of the drug is excreted in the urine within 8 hours after administration. The half-life in blood plasma is approximately 1.3 muscle hours. Table 1 Blood plasma concentration (ship/private land S.F.) 7-[D-Q-amino-Q-(p-hydroxyphenyl)-3-(1.2, 3-triazol-5-ylthiomethyl)-3-cephem -4-carboxylic acid propylene glycolate time (hr) 0.08 0.0
4±-0.25
0.3 Sat 0.10.50
5.9shi 2.00.75
8.4 eleven 1.0
15.8 Sat 3.21.5
17.7±2.0
2.0 18.
7±0.93.0
13.7±1.24.0
8.4±1.36.0
3.2±0.480
1.6±0.2 Additional relationship The invention of Rokusho No. 481144194 is "compounds having the D-(1) configuration in the side chain of the formula and easily cleavable esters thereof and salts usable as pharmaceuticals" in the preparation of a compound of formula 1 or a readily cleavable ester thereof or a salt thereof by reacting with an acylated derivative of formula and after removal of the amino protecting group a compound of formula 1 or a readily cleavable ester thereof or use as a medicament. If desired, before or after removal of the protecting group, (a) the free acid or the product in the form of its salt can be cleaved off correspondingly easily by methods known per se; (b) converting the product in the form of an ester or salt which can be easily cleaved by methods known per se into the corresponding free acid compound or into a pharmaceutically usable salt thereof; 2. A method for converting salts into salts which can be used as salts.

In addition, the invention of Tokugan Sho 48-1144194 includes 7-[D
A method for isolating -Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3 triazol-5-ylthiomethyl)-3-cefemu 4-carboxylic acid methanolate is disclosed.

In contrast, the present invention provides 171[D1Q-amino-Q
-(p-hydroxyphenyl)acetamide]-131(
1,2,3-triazol-5-ylthiomethyl)-3
-Cephemu 4-carboxylic acid 1,2-propylene glycolate, in which 7- [D-Q-
Amino-Q-(p-hydroxyphenylacetamide)
Increasing the pH of an acidic solution of 1-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid or its hydrate or solvate to 4.0 or higher by adding a base. Let me do it.

. and 27-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1, In the method for producing 2,3 triazol-5-ylthiomethyl)-13-cephem-4-carboxylic acid monopropylene glycolate (
11 7-[D-Q-amino-Q-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid or hydrate or solvate thereof Prepare an aqueous solution consisting of a water-lacquered organic compound having a compound and a ketone functional group; ■ Adjust the pH value of the solution to about 4.5; {3
1. Dilute the solution with a sufficient amount of water to evaporate water-based impurities; {4} Separate the aqueous solution from insoluble impurities; '5} Add a sufficient amount of 1,2-bromine to the aqueous solution. ``6} A method characterized by adding pyrene glycol to precipitate the target monopropylene glycolate; recovering a crystalline product.''

Therefore, the present invention is an invention in which the main part of the invention of Tokusho No. 48-144194 is an essential part of its structure, and it achieves the same object as the invention of Tokushi No. 48-144194. It is something to do.

Claims (1)

[Scope of Claims] 1 7-[D-α-amino-α(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carvone In the method for producing acid 1,2-propylene glycolate, 7-[D-α-amino-α-(p-hydroxyphenyl)acetamide] having a pH of about 2.0 or less dissolved in aqueous 1,2-propylene glycol. -3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4
- A process characterized in that the pH of the acidic solution of the carboxylic acid or its hydrate or solvate is raised to above 4.0 by addition of a base to precipitate the propylene glycolate product and then recover it. . 2 7-[D-α-amino-α-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid monopropylene glycolate In the manufacturing method of
(1) 7-[D-α-amino-α-(p-hydroxyphenyl)acetamide]-3-(1,2,3-triazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid or its prepare a water-soluble solution consisting of a hydrate or solvate and a water-soluble organic compound having a ketone functional group; (2) adjust the pH value of the solution to about 4.5; (3) make the solution sufficiently (4) Separate water-soluble from insoluble impurities; (5) Add a sufficient amount of 1,2-propylene glycol to the aqueous solution to obtain the desired monomer. - precipitating propylene glycolate; (6) collecting the crystalline product;